Biomedical Applications of On-Line Preconcentration-Capillary Electrophoresis Using an Analyte Concentrator: Investigation of Design Options

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Abstract
A method to perform on-line sample preconcentration of serum immunoglobulin E by affinity capture is described. Purified anti-IgE antibodies were covalently bound to an analyte concentrator-reaction chamber or cartridge. The immunoglobulins (IgE) were bound to and eluted from the cartridge by the optimum dissociating buffer system, and the eluent(s) were then subjected to capillary electrophoresis. The first design used was a 5 mm solid-phase cartridge fabricated by assembling a bundle of multiple microcapillaries in which a monoclonal antibody directed against IgE was covalently bound to the surface of every microcapillary. The whole assembly was connected, through sleeve connectors, to the capillary column for affinity capillary electrophoresis. The second design used consisted of an analyte concentrator-reaction chamber that was fabricated from a solid rod of glass. Several small diameter passages or through holes containing a similar surface area was tested for the same experiments and performance as described above. A major advantage of these designs, over previously described designs, is the absence of frits and beads. The previously reported designs consisted of derivatized beads confined to the concentrator cartridge by a frit at each end. After limited usage of the cartridge, the beads tends to pack at the outer frit. This lead to restricted flow through the concentrator chamber and ultimately clogging of the system. The designs reported here allows for a constant electroosmotic flow, superior reproducibility of the electropherograms, a reduced possibility of blocking the microcapillaries, and increase number of usages of the cartridge. The use of this novel analyte concentrator design for the determination of immunoglobulins in biological fluids is demonstrated by capillary electrophoresis of IgE in serum. The general utility of this technique for a variety of biomedical applications is discussed.

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